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BM28 is a phosphoprotein that migrates at 125kDa in SDS PAGE with a hyperphosphorylated form migrating as a slightly slower band. The ratio of the phosphorylation states alters with the phases of cell cycle - in M, the “fast” phosphorylated BM28 is the major form found, while in G1 phase, the “slow”, hyperphosphorylated BM28 predominates. The cellular localization of BM28 is also cell cycle dependent. In G1, most of the BM28 is chromatin bound, whereas, by M phase, the BM28 is still nuclear, but not associated with the DNA. BM28 is necessary for both entry into S phase and cell division as determined by microinjection inhibition experiments. Microinjection of a BM28 antibody into synchronised cells in G1 phase inhibits DNA replication. When injected during S phase or later, no effect on DNA replication is seen, but cell division is inhibited.Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Western Blotting
